Concentration and recovery of carotenoid pigments from palm oil



patented ()ct. 23, i951 UNITED STATES PATENT OFFICE 1 I v 2,572,467 v QCONCENTRATION AND RECOVERY OF CA- ao'rENom PIGMENTS FROM PALM 01L ArthurIra Gebhartg; Union, N. J., assig'nor to Qolgate-Palmoliv'e-PeetCompany, Jersey City,- N. J a corporation of Delaware No Drawing.Application August 4, 1949; Serial N0. 108,649

12 Claims. 1

This invention relates to an improved process for the concentration andrecovery of the caroteno-id pigments, and more specifically the'carovtenes or provitamins A from palm oil.

Any means of recovery of the carotenoid content and the like must takeinto consideration the complex character of the parent palm oil materialand of the carotenoid substances contained therein. Palm oil is amixture of triglycerides of a variety of higher fatty acids andpossesses generally significant amounts of free acids in addition tounsaponifiable matter such as the carotenoids, sterols, nitrogenousmatter, etc. The carotenoid pigments are present in relatively minutequantities of the order of about 0.05-0.20%; they are heat and airunstable, of high molecular weight, oil soluble and hydrocarbon incharacter generally. The aggregate of these various factors render theconcentration and recovery of the carotenoids diflicult in prior artprocesses.

Heretofore, a common technique for the recovery of carotenoid pigmentshas involved usually the saponification of the fatty oil and extractionof the unsaponifiable fraction with a suitable selective solvent. Thisprocedure is limited by the necessity for handling comparatively largevolumes of immiscible and frequently emulsifiable liquids and theprovitamin A fraction is recovered admixed with other unsaponifiablematter. In another process, crude palm oil may be dissolved in a solventand a certain portion of the glycerides and the free fatty acids may beseparated therefrom by crystallization. Thereafter, the residual fattyacids may be neutralized and a soap phase separated therefrom. Therecovery of the "carotenoid pigments is incomplete therein due to anaffinity or solubility of the triglycerides for the carotene substances,and to the possible formation of emulsion in the presence of the soapwhich would interfere with the sharp separation of an aqueous phase.

Another procedure different in character involves the alcoholysis ofpalm oil and the separation of the provitamins by distillation. Thisprocedure results in fair yields but involves the use of hightemperatures and complex and costly operating techniques including highvacuum distillation A simple and economical process has now beendeveloped for-the concentration and recovery of the carotenoid fractionfrom palm oil. Broadly, the present invention comprises theconcentration of the carotenoid pigments from carotenoid-.- c'ontainingfatty mixture derived from the by- 260-2265) 2 drolysis and alcoholysisof palm oil by treatment with an inert selective solvent andcrystallization of at least the major portion of the fatty productspresent in such mixtures therefrom. By means of the application ofvarious splitting processes, the original palm oil may be renderedsubstantially free of triglycerides and converted simultaneously tocompositions comprising essentially free fatty acids or their alkylesters, which are amenable to solvent crystallization at lowtemperatures of the fatty constituents such that the desired carotenoidcompounds may be recovered in a highly concentrated form and unimpairedcondition from the remaining solution.

Accordingly, the present invention contemplates the hydrolysis of thepalm oil in any suitable manner such that decomposition of thecarotenoid pigments does not occur appreciably. Since the carotenoidsubstances are heat labile, the conventional types of splittingprocesses utilizing high temperatures of the order of 250. C. areundesirable herein. A preferred manner of preparing a hydrolyzed palmoil involves the two stage process of saponification of the palm oil atrelatively low temperatures and acidification of the resulting soaps tothe freeacids.

The palm oil may be saponified in any suitable manner, either by batch,semi-continuous or continuous operations. The saponification isaccomplished by the reaction of the palm oil with an alkalinesaponifying agent or mixtures of such agents, preferably those having analkali metal or alkaline earth metal as the cation. Since relatively lowtemperatures are to be employed, it is preferred to use the causticalkalies such as sodium and potassium hydroxide. The proportion ofalkaline saponifying agent should be sufiicient to insure substantiallycomplete saponification of the fatty acidspresent in both the free stateand in combined form as esters in the palm oil. Accordingly,approximately a molar equiv-' alent and preferably an excess (generallyup to about a 25% excess) of saponifying agent may be utilized in orderto effect saponification. The saponifying agent may be added either asan aqueous solution or. dissolved ina suitable'diluent, particularly anorganic solvent such as ethyl alcohol.

During saponification, any appreciable decomposition of the carotenoidfraction should be avoided to insure maximum recovery of the same. Thesuitable temperature range should be less than about 150 C., andpreferably not more than 100 C. with best results attainable by the useof temperatures not greater than about C. If

desired, reduced pressure may be employed advantageously as a means oflimiting temperature and oxidation effects. Saponification with analcoholic caustic solution is a particularly satisfactory proceduresince the reaction may be conducted 'WlthQllt carotenoid decompositionat about the boiling point of the alcohol.

The time of heat treatment is always a factor to be considered andcorrelated with the particular temperature selected for the reaction. Ifappreciable decomposition of the carotenoids 'oc curs at the highertemperatures,'the temperature and/or time of treatment should'be reducedaceordingly;

At ordinary temperatures, the carotenoid pigments in palm oil arereasonably stable to air but characteristically are liable todecomposition by oxidation, particularly at elevated temperatures. Thepresence of air therefore at the temperatures to be employed during thesaponification reaction is not .p'erf-erred, though it need not beexcluded'necessarily. The use of a closed system, a -vacuum'or'of'an-inert atmosphere (e. g. nitrogen) may be employed advantageously.

The saponified mass substantially completely free of triglycerides maythen be subjected to acidification in any suitable manner to convert thesoaps to the free fatty acids. A common procedure involves the additionof a suitable acidic material, -e. g.-a dilute solution of sulphuric,phosphoric "or hydrochloric acids, in an amount sufficient to attain atleast about a pH of '7 and preferably about 4 or 5 in the reactionmixture.

Upon acidification and settling, two layers are formed which maybe'zseparated preferably. The lower aqueous layer contains the solublesalts, glycerine and excess acid and the upper fatty acid layer'containsthe carotenoids. If desired, this upper layer maybe water washed anddried to achieve additional purification before subjecting the mass tothe requisite solvent crystallization treatment.

Similarly, the palm oil may be subjected to alcoholysis to form thedesired readily crystallizable mixture. et al. U. S. Patent No.2,383,579; Dreger, U. S. Patent No. 2,383,596; and Trent, U. S. PatentsNos. 2,383,632 .and 2,432,181 are applicable herein.

In such alcoholysis processes, the palm oil is treated at lowtemperatures with a suitable monohydric alcohol, generally in thepresence of an alcoholysis catalyst whereby conversion of thetriglycerides to the fatty acid esters of the monohydric alcohols occursrapidly. The monohydric alcohols arepreferably. the saturated aliphatic01165701. not more than about 6 carbon atoms, such as-methyL'ethyl,propyl, butyl alcohols, etc.

The rate of the alcoholysis reaction is largely determined by thetemperature and the presence or absence of suitable catalysts. If acatalyst is used, an advantageous temperature range is from about 20 to150 'C. The alcoholysis reaction proceeds'q'uite rapidly even at roomtemperature in the presence of a catalyst but proceeds even more rapidlyat elevated temperatures. Suitable preferred catalysts are the alkalinematerials suchas sodium hydroxide, sodium alcoholate, etc. Since thecarotenoid compounds are heat unstable, the use of high temperaturestends to promote decomposition of the :same. In order to .preventsubstantial decomposition of the carotenoid pigments, the temperatureand the time of treatment should be kept to a minimum consistent withsubstantially complete conversion of the triglycerides to the alkylesters. Thus the The processes disclosed by Allen treated in anysuitable manner to remove unreacted alcohol, glycerine, etc. As apreferred technique, the reaction mixture may be allowed to settle intolayers wherein the upper layer contains the alkyl esters, excessalcohol, and unsa'pbnifiables including the carotenoids. and theglycerine is concentrated with some alcohol in the lower layer. The twophases may be separated in any convenient manner such as by decantation.The upper layer may be washed with water or aqueous alcohol to separateresidual glycerine and the excess alcohol. Alternative to settling, theentire reaction mass may be washed as-a means of removing the glycerineand excess alcohol or the alcohol could be removed by distillation. Thealkyl ester fraction containing the carotenoids may now be subjected tothe crystallization procedure. It is preferable however to neutralizethe alkaline catalyst, if one has been employed for the alcoholysisreaction, either before the washing or the crystallization treatments'to remove any .possible soap which has formed in the reaction mixture.

Alternative to alcoholysis as a means of .pre: paring the monoesters,there may be employed direct esterification of the free fatty acidsderived by hydrolysis, with :a suitable alcohol by heating aconcentrated mixture of the reactants, generally in the presence of anesterification catalyst.

Broadly, solvent crystallization as a means of purification of a desiredconstituent is an old procedure. In the fractional crystallization "ofany complexsystem, such as the mixtures specifiedherein, there areinvolved a numberof factors. Such a system exhibits the solubility ofthe individualcomponents in the particular solvent, and also the mutualsolubility of the substances upon one another. While the former factorcan be determined by actual solubilityexperiments upon the pureindividual .substances, the vlatter factor is more complex and involvessuch considerations as mixed crystal formation, association between thecomponents themselves or between the components and the solvent, etc.

If .palm oil itself were crystallized, sharp separation of thecarotenoid content therefrom would be difficu lt and incomplete sincethe carotenoid compounds exhibit a marked aflinity for thetrigly'cerides, and the triglycerides tend to assume the averagecharacteristics of the fatty acids as generally two or three differentacids of diverse nature and properties maybe combined within a singletriglyceride molecule. By the'conversion of the triglycerides to thefree fatty acids or their simple .alkyl esters, these latterconsiderations are-markedly reduced and eliminated for practicalpurposes.

Thus, it has been found that the carotenoidcontaining mixtures composedprincipally of the free :fatty acids or the alkyl esters and derived byhydrolysis .or alcoholysis respectively of the parent palm oil may Jaesubjected to solvent crystallizationto attain an unusually sharp separaealcohols, esters, the hydrocarbons, halogenated hydrocarbons, ethers,etc. More specific examples of fatty solvents suitable for fractionalcrystallization herein are acetone, diethyl ketone, propane, butane,hexane, heptane, petroleum ether, trichloroethylene, propylenedichloride, chloroform, methyl alcohol, ethyl alcohol, carbondisulphide, methyl acetate, ethyl acetate, isopropyl acetate, monoethylether of diethylene glycol, etc. The solvents may be strongly or weaklypolar, or non-polar in character. It is preferred however to use thenormally liquid solvents such as acetone, methyl and ethyl acetate, etc.and

preferably those having a boiling point of not more than about 100 C.

The amount of inert solvent used to dissolve the fatty acid or fattyacid monoester composition may vary somewhat depending upon theparticular selective solvent. The use of a concentration of thecomposition in the solvent of as low as about 2% is operable but aconcentration of about 10% to about is preferred, withparticularlysatisfactory results being attained within the range from about 10% toabout 20% concentration.

After dissolving the mixture derived from palm oil in the solvent, thesolution may be cooled to predetermined temperatures such thatsubstantially all the fatty acids or fatty acid monoesters presenttherein are precipitated as a solid phase. The mixture may be separatedby any means suitable for solid-liquid phase separation, includinggravity, filtering, centrifugal force etc. The liquid or filtrate mayagain be cooled to the same or lower temperatures and additionalsolidified fractions may be separated therefrom.

Any number of cooling stages may be employed; however it is preferred tocarry out at least two or three separation procedures. The separatedmatter if desired, may be further purified by dissolving it in freshand/or a different solvent and reprecipitating the solid matter fromsolution by cooling. In addition, the separated matter may be washed oneor more times with solvent at suitable temperatures.

In order to obtain a highly concentrated ca.- rotenoid fraction, thesolution should generally be cooled to at least about 40 C., andpreferably about -50 C. At these temperatures, substantially all theundesired constituents have been precipitated and separated thereby fromthe carotenoids in solution. Much unsaponifiable mat ter, substantiallyall the saturated fatty acids such as myristic, palmitic, and stearic,and a considerable percentage of the unsaturated fatty acids havecrystallized out of solution at these temperatures.

The residual solution or filtrate is generally orange in color andcontains substantially all the carotenoids in an unimpaired conditionadmixed with minor amounts of unsaturated acids. The unsaturatescomprise chiefly linoleic acid and a minor amount of oleic acid...If"..desired, this solution may be recooled at lowertemperaturesof'the orderxof about =50 C. and 70. C. whereupon yellow toorange-colored crystals are precipitated from solution. These crystalsare converted to an orange colored oil at room temperature. This orangecolored oil contains the carotene isomers as its chief constituents, thelargest proportion being beta carotene, then alpha carotene and smallamounts only of gamma carotene and other carotenoid pigments.'Minor'amounts of unsaturatedfatty acids'comprising chiefly linoleicassociated with a small proportion of oleic acid may be identified inthe oil. Upon standing, the color of the orange oil is rapidly bleachedby exposure to air and light to a very light greenish-yellow color. I

The solvent may be removed from the concentrate in any suitable manner.Where carotenoid crystals have been precipitated from solution, it isobvious that any conventional solid-liquid phase separationjmeans maybeemployed as indicated supra. Where the carotenoid pigments are insolution, the solvent may be removed by evaporation. The solution may beheated under vacuum and preferably while passing in an inert gas toprevent injury to the'carotenoid sub-- stances during distillationof thesolvent.

If desired, the various carotenoid substances in the concentrate may beisolated in any suitable manner. Fractional adsorption on alumi' numoxide or fullers earth may be used'satisfac torily. Chromatographicadsorption on calcium hydroxide or magnesium hydroxide from a solutionof the carotenoids in a suitable solvent after I removal of any fattyacids are additional methods of purification. I

The following are additional specific examples designed to beillustrative to the nature of the invention:

1 Example I Red palm oil is saponified by the addition o f 'a 10% excessof potassium hydroxide, added as a 20% solution of KOI-I in ethylalcohol, at 75-80? C.'in an atm-osphereof nitrogen. The saponificationreaction mixture is acidified with dilute jsulphuric acid to convert thepalm oil soaps to the corresponding free acids. After allowing thereaction mixture to settle, the aqueous phase/containing the excesssulphuric acid, glycerinefand alcohol is removed and the fatty acidphase is recovered for treatment.

50 grams of the palm oil acids are dissolved in two liters of acetoneand cooled to *I0 C. for 20 minutes whereupon a light coloredfractionprecipitates from solution. Themixture is then filtered toremove the solids, which are Washed with ml. of acetone previouslycooled to -10 C. The washed solids are then almost white in color andcomprise principally the more saturated fatty acids present in theoriginal palm oil. The acids are washed with ether and weigh 17.0

grams. 7

The filtrate possessing the original orange color is recooled to 55 C.and the. precipitated fatty acids are separated by filtration. Afterwashing with two 150 ml. portions of acetoneat 50 C, the solidifiedfatty acids are dried and weigh 22.0

grams. r

The crystallizataion procedure is repeated again on the filtrate at atemperature of -70 C. A small crop of yellow colored crystals separateswhich upon purification weighs 3.0 grams and is converted to an orangecolored oil at room temperature. Upon standing, the col-or bleachesrapidly to a very light greenish-yellow color. Analysis of a. sample ofthe orange colored oil 7 discloss'th carotene isomers to'b the chiefconstituents (the largest proportion'being beta caro ten'e, then .alphacarotene and small-amounts only of gammacarotene) Minor amounts ofunsaturated fatty acids comprising chiefly linoleic .acid associatedwith a smallproportion of oleic .acid may be identified in theconcentrate.

Example II Red palm oil issaponified with a 10% excess of 50 .Baumaqueoussodium hydroxide solution at 80 C. under a reduced pressure-offifteen inches of mercury in an atmosphere of nitrogen. Thesaponification reaction mixture is acidified with dilute sulphuricacid-and the resulting fatty acid phase containing the carotenoids isseparated as indicated in Example I. i

The fatty acid layer is dissol-ved in sumcient petroleum ether to form a10% solution. The solvent solution is cooledslowly to -C. and asolidified fraction which separates out-isremoved by filtration. Theprocedure is repeated upon the filtrateat C., and at C. The finalfiltrate has a deep-orange color and-the-petroleum ether solventis-removed by evaporation. A sample of' the orange coloredsolvent-freeliquid is subjected to chromatographic adsorption and indicates that thecarotenoid content consists chiefly of the :alpha and'beta caroteneisomers, with. minor amounts of other carotenoid pigments. 1 V

. ExampleIII "Palm oil is subjected to alcoholysis at 45 C. withinethanol --using 15 moles of .methanol per mole of palm oil in thepresence of sodium hydroxide as a catalys't. At the completion of thereaction, "the alkaline catalyst is neutralized by the addition ofdilute sulphuric acid. The reaction mixture is allowed to separate intotwo layers with .the-upperelayer containing the alkyl esters, alcohol,and the .unsaponifiablesincluding the carotenoids, and the lower layercomprising the glycerine and alcohol. The lower layer is removed and-theupper ester layer is washed and dissolved in sufficient petroleum etherto forma 20% solution.

The petroleum ether solution issubjeted to fractional crystallization at10 C., 20 -C., an'd C. in the manner indicated in Example 1 whereby thesaturated "fatty acidmonoesters-and the vast proportion of theunsaturated fatty acid monoesters are removed .by filtration. Thesolvent filtrate remaining after removing the fraction at minus 50 0.possesses an .orange color and is al'iighly concentrated carotenoidlfraction con- "taining minor amounts of methyl esters ofunsaturatedfatty acid. The solvent'is evaporated under vacuum'andthecarotenoid concentrate is quickly placed in .a containerin an atmosphereof'nitrogen to prevent oxidationa'ndloss of caroterm. A sample of theconcentrate is subjected to adsorption upon alumina whereby the carotenecontent .is substantially separated fromtheresidual" fatty acidmonoesters, and 'dis'clo'singthereby in addition thepresence ofcomparatively "large amountsof alpha and betacarotenes.

Example IV The ethyl esters of :palm oil acids =arejpre- ;pared in the'manner indicated -in Example .11 1 "using a 15 ;to 1 molar ratiozof.alcohol toijpalm -oil.- Upon completion of :the reaction ;and-'set--tling, approximately .4 .molesof ethanol .per;m ole .of ethyl estersremain inrthe-upper layer of the reaction mixture.

The upper layer containing .the ethyl esters, the carotenoids and excess:a'lcohol :is subjected tofractional crystallization at -l0 -25" and 45"C. using the excess :ethyl alcohol as a solvent and diluent. The majorproportion of the ethyl esters areprecipitated therebyizisolid ifiedform. After separating the precipitate. by filtration,- a finalsolidified orange-red fraction is crystallized from the filtrate at C.and comprises chiefly the desired carotenoid constituents admixture witha minor amount of unsaturated fatty acid ethyl esters.

Since certain changeszmay beimadein carryi out the above process withoutdeparting from the scope of the invention, it is intended that all:matter contained in the above description shall .be interpreted asillustrative and not in a limiting sense.

It is also to 'be understood that the following claims .are intended tocover all the eneric and specific features of the invention .hereindescribed, and all statements of the SCODBOfqthB invention, which as amatter of language might be said to fall therebetween.

Having described the invention, what is claimed as new and desired to besecured by Letters Patent is:

1. The process of recovering a carotenoidaconcentrate from .acarotenoid-containing composition comprising chiefly .a fatty materialof 'the group consisting of palm oil fatty acids and fatty acidmonoesters of lower monohydric alcohols substantially free oftriglycerides derived from the hydrolysis and alcoholysis of "palm .oilrespectively, which comprises dissolvingsaidcomposition in an inertorganic solvent, .coolingthe resultant solution to precipitate therefromat least the major portion of the fatty material, separating theprecipitated fraction vandrecovering .a carotenoid concentrate.

2. The process for preparing a carotenoid concentrate from acarotenoid-containing fatty composition of the group consisting of fattyacid mixtures and lower alkyl fatty acidmonoester mixtures derived fromthe hydrolysis and a1 coholysis of palm oil respectively, which,comprises cooling a solution of .thesame in an inert organic solvent toat leastabout 40 C..to crystallize the majorpor-tion of the'fattyconstituents therefrom, and separating the crystallized material "fromthe remainingcarotenoid-containing solution.

3. The process of claim 2 wherein the solvent is a normally liquidorganic solvent.

4. The process of concentratingthe carotenoid content of palm oil whichcomprises ."convert ing said palm oil to a ,fatty material containingprincipally afatty constituent of the. group ,consisting of fatty acids:by hydrolysis and lower alkyl fatty :acid monoesters .by alcoholysisrespectively admixed with the carotenoid c onstituents and substantiallyfree of triglycerides dissolving :said fattymaterial in anzinert organic:solvent to form a solution, :lowering the temperature-of the resultingsolution to crystallize at leastxarmajorportion of the-fattyconstituentstherefrom, separating saidcrystallizedzfattyaconstituents and recoveringa carotenoid .aconcentrate from fthe residual solution.

'5. *The process -.of claims wherein the solvent is a normally liquidorganicisolvent.

i6. Thepro'c'ess of :re'coveringa carotenoid fraction 'from palm oilwhich comprises :subjecting said palm oil toalcoholysiswith=ailowerzmonohydric' .alcohol "to. produce '23,creactionza-mixture containing chiefiy fatty acid esters of saidmonohydric alcohol admixed with the carotenoid substances, dissolvingsaid fatty acid monoe'sters and carotenoids in an inert organic solvent,lowering the temperature of the resulting solution to a predeterminedpoint to cause solidification of at least the major portion of saidfatty acid monoesters, separating the solidified fraction from theremaining solution, and recovering a highly concentrated carotenoidfraction.

7. The process of recovering a carotenoid concentrate from palm oilwhich comprises subjecting said palm oil to alcoholysis with a lowermonohydric alcohol to produce a reaction mixture containing chieflyfatty acid esters of said monohydric alcohol admixed with the carotenoidsubstances and substantially free of triglycerides, dissolving saidfatty acid monoesters and carotenoids in an inert organic solvent,lowering the temperature to at least minus 40 C. to precipitate asolidified fraction, separating the solidified fraction, and recoveringa highly concentrated carotenoid fraction from the remaining solution.

8. A method for the recovery of a carotenoid concentrate from palm oilcomprising alcoholizing said palm oil with a lower monohydric alcohol toproduce a reaction mixture containing principally fatty acid monoestersadmixed with the carotenoids, subjecting said mixture to fractionalcrystallization with an inert organic solvent to remove the majorportion of the fatty acid monoesters, and recovering a carotenoidconcentrate.

9. A method for the recovery of a carotenoid concentrate from palm oilcomprising hydrolyzing said palm oil, to produce a reaction mixturecontaining principally fatty acids admixed with the carotenoids,subjecting said mixture to fractional crystallization with an inertorganic solvent to separate the major portion of the fatty acids fromthe carotenoids and recovering a carotenoid concentrate thereby.

10. A method for the recovery of a carotenoid concentrate from palm oilcomprising hydrolyzing said palm oil by saponification and acidificationto produce a reaction mixture containing principally fatty acids admixedwith the carotenoids, subjecting said mixture to fractionalcrystallization with an inert organic solvent to separate the majorportion of the fatty acids from the carotenoids, and recovering acarotenoid concentrate.

11. A process for recovering a carotenoid concentrate from palm oilwhich comprises saponi fying said palm oil with an alkaline saponifyingagent at a temperature of less than about C. to convert triglycerides ofpalm oil to soaps, acidifying said soaps to free fatty acids containingcarotenoid pigments of palm oil in admixture therewith, dissolving theresulting fatty acidcarotenoid mixture in an inert organic solvent,lowering the temperature of said mixture to crystallize at least a majorportion of the fatty acids therefrom, separating said crystallized fattyacids and recovering a carotenoid concentrate.

12. The process of claim 11 wherein fatty acids are crystallized attemperatures of at least -40 C.

ARTHUR IRA GEBHART.

REFERENCES CITED The following references are of record in the file ofthis patent:

UNITED STATES PATENTS Number Name Date 2,318,747 Buxton May 11, 19432,460,796 Eckey Feb. 1, 1949 FOREIGN PATENTS Number Country Date 433,930Great Britain Aug. 22, 1935

11. A PROCESS FOR RECOVERING A CAROTENOID CONCENTRATE FROM PALM OILWHICH COMPRISES SAPONIFYING SAID PALM OIL WITH AN ALKALINE SAPONIFYINGAGENT AT A TEMPERATURE OF LESS THAN ABOUT 150* C. TO CONVERTTRIGLYCERIDES OF PALM OIL TO SOAPS, ACIDIFYING SAID SOAPS TO FREE FATTYACIDS CONTAINING CAROTENOID PIGMENTS OF PALM OIL IN ADMIXTURE THEREWITH,DISSOLVING THE RESULTING FATTY ACIDCAROTENOID MIXTURE IN AN INERTORGANIC SOLVENT, LOWERING THE TEMPERATURTE OF SAID MIXTURE TOCRYSTALLIZE AT LEAST A MAJOR PORTION OF THE FATTY ACIDS THEREFROM,SEPARATING SAID CRYSTALLIZED FATTY ACIDS AND RECOVERING A CAROTENOIDCONCENTRATE.